Ccl5 inhibitors

ABSTRACT

Compounds, pharmaceutically acceptable salts, esters, prodrugs, and pharmaceutical compositions thereof are disclosed that are useful for inhibition of the biological activity of CCL5 on mammalian cells, as well as methods of treatment for diseases involving the increased biological activity of CCL5.

FIELD OF THE INVENTION

This invention pertains to inhibitors of the biological effects of CCL5on mammalian cells. The invention provides compounds and methods usefulfor inhibition of the biological effects of CCL5 on mammalian cells andfor the treatment of dermatologic, ocular, respiratory, and otherdiseases associated with increased CCL5 biological activity on humancells and tissues.

BACKGROUND OF THE INVENTION

Chemokines constitute a family of chemoattractant cytokines and aresubdivided into four families on the basis of the number and spacing ofthe conserved cysteine residues in the N-terminus of the protein.Chemokines play a major role in selectively recruiting monocytes,neutrophils, and lymphocytes, as well as in inducing chemotaxis throughthe activation of G-protein-coupled receptors. CCL5 (also known asRANTES) belongs to the C—C chemokine subfamily, whose chemokines presentadjacent cysteines, and comprises the majority of the chemokines [1, 2].This chemokine plays an important role in leukocyte biology and diseasepathogenesis by controlling cell recruitment and activation in basal andin inflammatory circumstances. In addition, because chemokine receptorsare expressed on other cell types, chemokines have multiple other roles,including angiogenesis, tissue and vascular remodeling, pathogenelimination, antigen presentation, leukocyte activation and survival,chronic inflammation, tissue repair/healing, fibrosis, embryogenesis andtumorigenesis [3,4]. The many biologic effects of chemokines aremediated by their interaction with chemokine receptors on the cellsurface. The most relevant known receptors for CCL5 are CCR1, CCR3, andCCR5 [I].

CCL5 has been shown to induce the in vitro migration and recruitment ofT cells, dendritic cells, eosinophils, NK cells, mast cells, andbasophils. Although initially considered to be a T cell-specificcytokine (hence the original name, RANTES; regulated upon activation,normal T cell expressed and secreted) [5], CCL5 is produced byplatelets, macrophages, eosinophils, fibroblasts, endothelium,epithelial, and endometrial cells. The variety of cells that express andmediate CCL5 effects implicates this chemokine in multiple biologicalprocesses, from pathogen control to enhancement of inflammation inseveral disorders, such as: arthritis, asthma, cancer, dermatitis,endometriosis, lichen planus, psoriasis, Sjogren syndrome, uveitis, andothers [6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16].

Certain types of normal mammalian and human cells such as; endothelial,epithelial, fibroblast, keratinocyte, smooth muscle, mesangial,astrocyte, monocyte, and microglial cells can be induced to produce andsecrete high levels of CCL5 by stimulation with various substances andunder a variety of stress conditions. In the resting state of thesecells the level of CCL5 is barely detectable. However, exposure of thesecells to various stimulants alone or in combination, such as cytokines;IL-1b, and TNF-alpha, cause increase in production of CCL5 [17]. Whenthese types of cells are subjected to stress conditions such as lowoxygen levels (hypoxia) they respond by producing increased amounts ofCCL5 [18]. In addition, when these types of cells are exposed to variousbacterial antigens or components of bacterial cell breakdown such aslipotechoic acid, flagellin, and lipopolysaccharide (LPS) they areinduced to produce and secrete increased amount of CCL5. In addition,substances that mimic bacterial, viral and fungal components such asPoly(I:C) and Pam3Cys also induce these types of mammalian cells toproduce increased amounts of CCL5 [19].

CCL5 binds to and transduces signals through the G-protein-coupledreceptors, such as; CCR5, CCR3, and CCR4. These receptors are present onmany types of human cells such as: leukocytes, peripheral bloodmononuclear cells (PBMCs), fibroblasts, vascular smooth muscle cells,endothelial cells, and astrocytes [20].

When CCL5 binds to its cell surface receptors on leukocytes it causesthem to produce cytokines such as; IL-1 alpha (Interleukin 1 alpha),IL-1 beta (Interleukin 1 beta), IFN-gamma (Interferon gamma), CCL2(chemokine motif ligand 2), and IL-8 (Interleukin 8) [21], whichparticipate in increasing inflammatory response and contribute to tissuedamage, pain, and loss of function in several inflammatory diseases.Binding of CCL5 to its receptors on endothelial cells causes them toproduce VEGF-A, an angiogenic protein.

Increased CCL5 production by human endothelial, epithelial, fibroblasts,keratinocytes, smooth muscle, mesangial, astrocytic, monocytic, andmicroglial cells promotes inflammation, inflammation-dependent diseaseprogression, subsequent tissue destruction, and loss of function invarious human tissues such as the dermis, epidermis, ocular uvea, andpulmonary bronchial epithelium by serving to attract monocytes,T-lymphocytes, and natural killer cells, and by increasing theproduction of pro-fibrosis genes in fibroblasts. Increased production ofCCL5 by cells and tissues leads to increased blood levels of CCL5. Aninherited genetic variant in the CCL5 gene promotor region is associatedwith several diseases [22, 23]. Therefore, CCL5 is an importanttherapeutic target to treat inflammatory diseases such as thosereferenced herein. There are no approved drugs on the market that reducecellular production of CCL5 [24]. Thus, there is a need for chemicalcompounds that inhibit the production of CCL5 by mammalian cells,especially those in epithelial tissues such as; skin, lung,gastrointestinal tract, eye, genito-urinary tract and endocrine glands,as well as in endothelial tissues.

SUMMARY OF THE INVENTION

According to one embodiment of the present invention, there is provideda compound of Formula 1 below:

wherein:n and p are independently selected from 0 or 1;R₁ and R₂ are independently selected from H, OH, F, Cl, Br, I,(halogen)alkyl, C₁-C₈ straight or branched chain alkyl, C₁-C₈cycloalkyl, heterocycloalkyl, alkylheterocycloalkyl, optionallysubstituted C₁-C₈ alkenyl, optionally substituted C₁-C₈ alkynyl,optionally substituted aryl, optionally substituted alkylaryl,optionally substituted heteroaryl, optionally substitutedalkylheteroaryl, O-alkyl, O-cycloalkyl, O-alkylcycloalkyl, O-aryl,0-optionally substituted aryl, alkyl-O-aryl, alkyl-O-optionallysubstituted aryl, C(O)-aryl, C(O)-optionally substituted aryl,CH₂C(O)-aryl, CH₂C(O)-optionally substituted aryl, 0-(halogen)alkyl;R₃, R₄, R₅, R₆, R₇, and R₈, if present, are independently selected fromH, C₁-C₈ straight or branched chain alkyl, C₁-C₈ cycloalkyl,heterocycloalkyl, alkylheterocycloalkyl, optionally substituted C₁-C₈alkenyl, optionally substituted C₁-C₈ alkynyl, optionally substitutedaryl, optionally substituted alkylaryl, optionally substitutedheteroaryl, or optionally substituted alkylheteroaryl, and/or adjacentsubstituents R₁ and R₂, R₃ and R₄, R₅ and R₆, R₇ and R₈, if present, mayform a saturated or unsaturated 3-7 membered carbocyclic or heterocyclicring; R₉ is selected from COOH, COO-straight or branched chain alkyl,B(OH)₂, B(OR₁₀)(OR₁₁);R₁₀ and R₁₁, if present, are independently selected from optionallysubstituted alkyl, cycloalkyl, alkylcycloalkyl, and/or when R₁₀ and R₁₁are present and adjacent to each other can together form an alkylbridged 5 or 6 membered heterocyclic ring, or a pharmaceuticallyacceptable salt, ester or prodrug form thereof with the proviso thatwhen n and p are both 0, R₁ is H, R₃ is methyl, R₄ is methyl, and R₉ isCOOH, COOCH₃, or COOC₂H₅, then R₂ cannot be C₁, and when and p are both0, R₂ is H, R₃ is methyl, R₄ is methyl, and R₉ COOH, COOCH₃, or COOC₂H₅,then R₁ cannot be C₁.

Additional embodiments of the invention include a pharmaceuticalcomposition comprising a compound as defined above and apharmaceutically acceptable carrier and/or diluent for topicalapplication.

Another embodiment of the invention is the inhibition of the productionof CCL5 by cells comprising mammalian epithelial tissues by topicalapplication of a compound or compounds according to Formula 1, eitheralone or in combination with other bioactive substances.

Yet another embodiment of the invention is the inhibition of theproduction of CCL5 by cells comprising mammalian vascular endothelialtissues by application of a compound or compounds according to Formula1, either alone or in combination with other bioactive substances, bydirectly contacting the endothelial cells.

An additional embodiment of the invention is the inhibition of thebiological effect of CCL5 on mammalian cells such as, but not limitedto, epithelial cells, monocytic cells, and endothelial cells, as well astissues containing these cells, by application of a compound orcompounds according to Formula 1, either alone or in combination withother bioactive substances.

A further embodiment of the invention is the use of a compound asdefined above in the manufacture of a medicament for the treatment of amammal at risk for or having at least one disease or disorder associatedwith elevated CCL5 levels. Non-limiting examples of such disorders are:inflammatory skin disorders and/or diseases; urticarial conditions;respiratory ailments; airway and pulmonary conditions; gastrointestinaldisorders; genito-urinary disorders; allergic diseases; atopicdisorders; infection-based diseases; trauma and tissue injury-basedconditions; fibrotic diseases; and/or ophthalmic/ocular diseases.

BRIEF DESCRIPTIONS OF THE DRAWINGS

FIG. 1a-b : Reduced production of CCL5 by human keratinocyte cells aftertreatment with compounds of Formula 1. FIG. 1a : cells were stimulatedwith Poly (I:C). FIG. 1b : cells were stimulated with Flagellin.

FIG. 2a-b : Decreased expression of the CCL5 gene in human keratinocytecells and human bronchial epithelial cells after treatment withcompounds of Formula 1. FIG. 2a : HEHK cells were stimulated with Poly(I:C). FIG. 2b : NHBE cells were stimulated with TNF-alpha.

FIG. 3: Reduced production of CCL5 induced by stimulation of humankeratinocyte cells with TNF-alpha after treatment with compounds ofFormula 1.

FIG. 4: Reduced production of CCL5 induced by stimulation of humanumbilical vein endothelial cells (HUVEC) with TNF-alpha after treatmentwith compounds of Formula 1.

FIG. 5a-d : Reduced production of IL-8 by human peripheral bloodmonocyte cells (PBMCs) and reduced production of VEGF-A by humanumbilical vein endothelial cells (HUVEC) induced by stimulation withrecombinant human CCL5/RANTES after treatment with compounds ofFormula 1. FIG. 5a : IL-8 gene expression. FIG. 5b : IL-8 Supernate.FIG. 5c : VEGF-A gene expression. FIG. 5d : VEGF-A Supernate.

DETAILED DESCRIPTION OF THE INVENTION

Compounds embodied by Formula 1 may have one or several asymmetriccenters and therefore can exist in different stereoisomericconfigurations. Consequently, the compound of Formula 1 can occur asindividual (pure) enantiomers, individual pure enantiomericdiastereomers as well as a mixture of enantiomers or diastereomers. Thescope of the present invention includes both single enantiomers andmixtures thereof in all ratios. The scope of the present inventionfurther includes all tautomeric forms (“tautomers”) of the compounds ofFormula 1, and all mixtures thereof in any ratio. It will be appreciatedby one skilled in the art that a single compound may exhibit more thanone type of isomerism.

The enantiomeric compounds of Formula 1 may be resolved into their pureenantiomers by methods known to those skilled in the art, for example byformation of diastereoisomeric salts which may be separated, forexample, by crystallization; formation of diastereoisomeric derivativesor complexes which may be separated, for example, by crystallization,gas-liquid or liquid chromatography; selective reaction of oneenantiomer with an enantiomer-specific reagent, for example enzymaticesterification; or gas-liquid or liquid chromatography in a chiralenvironment, for example on a chiral support with a bound chiral ligandor in the presence of a chiral solvent. It will be appreciated thatwhere the desired stereoisomer is converted into another chemical entityby one of the separation procedures described above, a further step isrequired to liberate the desired enantiomeric form. Alternatively,specific stereoisomers maybe synthesized by using an optically activestarting material, by asymmetric synthesis using optically activereagents, substrates, catalysts or solvents, or by converting onestereoisomer into the other by asymmetric transformation or inversion.

The compounds of the present invention may exist in unsolvated as wellas a variety of solvated forms with pharmaceutically acceptable solventssuch as water, ethanol, and the like. In general, the solvated forms areconsidered equivalent to the unsolvated forms for the purposes of thepresent invention. It should be understood that pharmaceuticallyacceptable solvents further include isotopically substituted solventssuch as D20, dimethyl sulfoxide-d6, and the like. The term ‘solvate’ isused herein to describe a complex comprising the compound of theinvention and one or more pharmaceutically acceptable solvent molecules,including water. As such, all manner of hydrates of the compound areincluded by the term ‘solvate’. It is intended that the presentinvention embrace unsolvated forms, solvated forms and mixtures ofsolvated forms in any ratio.

The compound of the present invention and/or its salts and/or solvatemay exist as amorphous solids or may exist in one or more crystallinestates, i.e. polymorphs. Polymorphs of the compound of Formula 1 areencompassed in the present invention and may be prepared bycrystallization under a number of different conditions such as, forexample, using different solvents or different solvent mixtures;crystallization at different temperatures; and using various modes ofcooling ranging from very fast to very slow during crystallization.Polymorphs may also be obtained by heating or melting a compound ofFormula 1 followed by gradual or fast cooling. The presence ofpolymorphs may be determined by solid NMR spectroscopy, IR spectroscopy,differential scanning calorimetry, powder x-ray diffraction or othertechniques. It should be understood that all such crystalline andamorphous forms of the compound of Formula 1, and its salts, solvates,and prodrugs thereof are encompassed by the invention and the claims.

The present invention also includes all pharmaceutically acceptableisotopically labeled variations of the compound of Formula 1. Suchisotopically labeled variations are compounds having the same Formulaand molecular formula as the compound of Formula 1 but wherein one ormore atoms are replaced by atoms having an atomic mass or mass numberdifferent from the atomic mass or mass number usually found in nature.Examples of isotopes that may be incorporated into the compound of thepresent invention include isotopes of hydrogen, carbon, fluorine,nitrogen, and oxygen, such as ²H, ³H, ¹¹C, ¹³C, ¹⁴C, ¹⁸F, ¹³N, ¹⁵N, ¹⁷O,and ¹⁸O, respectively.

Certain isotopically labeled variations of the compound of the presentinvention such as, for example, those incorporating a radioactiveisotope such as ³H and ¹⁴C, are useful in drug and/or substrate tissuedistribution studies. Tritium, i.e. ³H, and carbon-14, i.e. ¹⁴C, areparticularly preferred due their ease of preparation and detection.Further, substitution with heavier isotopes such as deuterium, i.e. ²H,can afford certain therapeutic advantages resulting from greatermetabolic stability, for example, increased in vivo half-life or reduceddosage requirements, and hence may be preferred in some circumstances.Isotopically labeled compounds of Formula 1 of this invention andprodrugs thereof can generally be prepared by carrying out theprocedures disclosed in the Schemes and/or in the Examples bysubstituting a readily available isotopically labeled reagent for anon-isotopically labeled reagent.

“Alkyl” means a straight or branched chain, saturated hydrocarbonradical. By way of example, the hydrocarbon chain may have from one totwenty carbons, one to sixteen carbons, one to fourteen carbons, one totwelve carbons, one to ten carbons, one to eight carbons, one to sixcarbons, one to four carbons, etc. “Lower alkyl” may refer to alkylshaving, e.g., one to six carbons, one to four carbons, etc. In certainexamples, a straight chain alkyl may have from one to six carbon atomsand a branched alkyl three to six carbon atoms, e.g., methyl, ethyl,propyl, 2-propyl, butyl (including all isomeric forms), pentyl(including all isomeric forms), and the like. “Me” means methyl, “Et”means ethyl, and “iPr” means isopropyl. Alkyl may be optionallysubstituted, e.g., optionally substituted with oxygen, silicon, sulphuror optionally substituted with OH, O-alkyl, SH, S-alkyl, NH₂, and/orNH-alkyl. In another example, alkyl may be C₁ to C₁₂ straight chain orbranched chain alkyl optionally substituted with oxygen, silicon,sulphur, or optionally substituted with OH, O-alkyl, SH, S-alkyl, NH₂,and/or NH-alkyl.

“Alkylene” means a divalent alkyl, with alkyl as defined above.

“Aryl” means a monocyclic or bicyclic aromatic hydrocarbon radical,e.g., having from 6 to 20 or 6 to 10 ring atoms e.g., phenyl ornaphthyl. Aryl may be optionally substituted, e.g., substituted phenylor substituted naphthyl.

“Alkylaryl” means a (alkylene)-R radical where R is aryl as definedabove. Alkylaryl may be optionally substituted. In certain examples,alkylaryl may be alkylphenyl, alkylsubstituted phenyl, alkylnaphthyl oralkylsubstituted naphthyl.

“Alkenyl” means a straight or branched chain, saturated hydrocarbonradical which contains a carbon-carbon double bond. By way of example,the hydrocarbon chain may have from two to twenty carbons, two tosixteen carbons, two to fourteen carbons, two to twelve carbons, two toten carbons, two to eight carbons, two to six carbons, two to fourcarbons, etc. “Lower alkenyl” may refer to alkenyls having, e.g., two tosix carbons, two to four carbons, etc. In certain examples, a straightchain alkenyl may have from two to six carbon atoms and a branched alkylthree to six carbon atoms, e.g., a vinyl group, an allyl group, butene(including all isomeric forms), pentene (including all isomeric forms),and the like. Alkenyl may be optionally substituted. In certainexamples, alkenyl may be a C₂ to C₁₂ straight chain or branched chainhydrocarbon containing a carbon-carbon double bond, optionallysubstituted with oxygen, silicon or sulphur, or optionally substitutedwith OH, O-alkyl, SH, S-alkyl, NH₂, or NH-alkyl.

“Alkynyl” means a straight or branched chain, saturated hydrocarbonradical which contains a carbon-carbon triple bond. By way of example,the hydrocarbon chain may have from two to twenty carbons, two tosixteen carbons, two to fourteen carbons, two to twelve carbons, two toten carbons, two to eight carbons, two to six carbons, two to fourcarbons, etc. “Lower alkynyl” may refer to alkynyls having, e.g., two tosix carbons, two to four carbons, etc. In certain examples, a straightchain alkynyl may have from two to six carbon atoms and a branched alkylthree to six carbon atoms, e.g., an acetylene group, a propargyl group,butyne (including all isomeric forms), pentyne (including all isomericforms), and the like. Alkynyl may be optionally substituted. In certainexamples, alkynyl may be a C₂ to C₁₂ straight chain or branched chainhydrocarbon containing a carbon-carbon triple bond, optionallysubstituted with oxygen, silicon or sulphur, or optionally substitutedwith OH, O-alkyl, SH, S-alkyl, NH₂, or NH-alkyl.

“Cycloalkyl” means a cyclic saturated or partially saturated hydrocarbonradical (or an alicyclic radical). By way of example, the cycloalkyl mayhave from three to twenty carbon atoms, from three to sixteen carbonatoms, from three to fourteen carbon atoms, from three to twelve carbonatoms, from three to ten carbon atoms, from three to eight carbon atoms,from three to seven carbon atoms, from three to six carbon atoms, etc.,wherein one or two carbon atoms may be replaced by an oxo group, e.g.,admantanyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,cyclohexenyl, indanyl, and the like.

“Alkylcycloalkyl” means a (alkylene)-R radical where R is cycloalkyl asdefined above; e.g., cyclopropylmethyl, cyclobutylmethyl,cyclopentylethyl, or cyclohexylmethyl, and the like. In another example,alkylcycloalkyl has four to twelve carbon atoms, i.e., C₄-C₁₂alkylcycloalkyl.

“O-alkyl” or “Alkoxy” means an (oxygen)-R radical where R is alkyl asdefined above. For example, O-alkyl may be an oxygen atom bonded to a C₁to C₆ straight chain or branched chain alkyl. Alkoxy groups have thegeneral formula: R-O.

“O-cycloalkyl” means an (oxygen)-R radical where R is cycloalkyl asdefined above. For example, O-cycloalkyl is an oxygen atom bonded to aC₃ to C₇ cycloalkyl.

“O-alkylcycloalkyl” means an (oxygen)-R radical where R isalkylcycloalkyl as defined above. For example, O-cycloalkyl is an oxygenatom bonded to a C₄ to C₈ alkylcycloalkyl.

“Heterocyclyl” or “heterocycloalkyl” means a saturated or unsaturatedmonocyclic group, in which one or two ring atoms are heteroatom selectedfrom N, O, or S, the remaining ring atoms being C. Heterocyclyl andheterocycloalkyl includes, e.g., where the heterocycle comprises one ortwo hetero atoms selected from O, S, or N, including a C₂ to C₆heterocycloalkyl. The heterocyclyl ring is optionally fused to a (one)aryl or heteroaryl ring as defined herein. The heterocyclyl ring fusedto monocyclic aryl or heteroaryl ring is also referred to in thisApplication as “bicyclic heterocyclyl” ring. Additionally, one or tworing carbon atoms in the heterocyclyl ring can optionally be replaced bya —CO— group. More specifically the term heterocyclyl includes, but isnot limited to, pyrrolidino, piperidino, homopiperidino,2-oxopyrrolidinyl, 2-oxopiperidinyl, morpholino, piperazino,tetrahydropyranyl, thiomorpholino, and the like. When the heterocyclylring is unsaturated it can contain one or two ring double bonds. Whenthe heterocyclyl group contains at least one nitrogen atom, it is alsoreferred to herein as heterocycloamino and is a subset of theheterocyclyl group. When the heterocyclyl group is a saturated ring andis not fused to aryl or heteroaryl ring as stated above, it is alsoreferred to herein as saturated monocyclic heterocyclyl.

“Alkylheterocycloalkyl” means an -(alkylene)-R radical where R isheterocyclyl ring as defined above e.g., tetraydrofuranylmethyl,piperazinylmethyl, morpholinylethyl, and the like. Alkylheterocycloalkylalso includes, e.g., where the heterocycle comprises one or two heteroatoms selected from O, S, or N and has three to eleven carbon atoms,i.e., C₃ to C₁₁ alkylheterocycloalkyl, and includes when N is present inthe heterocyclic ring the nitrogen atom may be in the form of an amide,carbamate, or urea.

“Heteroaryl” means a monocyclic or bicyclic aromatic radical, where oneor more, preferably one, two, or three, ring atoms are heteroatomselected from N, O, or S, the remaining ring atoms being carbon.Representative examples include, but are not limited to, pyrrolyl,thienyl (thiophenyl), thiazolyl, imidazolyl, furanyl, indolyl,isoindolyl, oxazolyl, isoxazolyl, diazolyl, pyrazolyl, triazolyl,benzothiazolyl, benzoxazolyl, quinolinyl, isoquinolinyl, pyridinyl(pyridyl), pyrimidinyl, pyrazinyl, pyridazinyl, tetrazolyl, and thelike. Heteroaryl may be optionally substituted.

“Oxo” or “carbonyl” means a=(0) group or C═O group, respectively.

The term “substituted” means that the referenced group is substitutedwith one or more additional group(s) individually and independentlyselected from groups described herein. In some embodiments, an optionalsubstituent is selected from oxo, halogen, —CN, —NH₂, —OH, —NH(CH₃),—N(CH₃)₂, alkyl (including straight chain, branched and/or unsaturatedalkyl), substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, fluoroalkyl, substituted orunsubstituted heteroalkyl, substituted or unsubstituted alkoxy,fluoroalkoxy, —S— alkyl, —S(O)₂-alkyl, —CONH ((substituted orunsubstituted alkyl) or (substituted or unsubstituted phenyl)), —CON(Hor alkyl)₂, —OCON (substituted or unsubstituted alkyl)₂, —NHCONH((substituted or unsubstituted alkyl) or (substituted or unsubstitutedphenyl)), -NHCOalkyl, —N(substituted or unsubstituted alkyl) CO(substituted or unsubstituted alkyl), —NHCOO (substituted orunsubstituted alkyl), —C(OH)(substituted or unsubstituted alkyl)₂, and—C(NH₂)(substituted or unsubstituted alkyl)₂. In some embodiments, byway of example, an optional substituent is selected from oxo, fluorine,chlorine, bromine, iodine, —CN, —NH₂, —OH, —NH(CH₃), —N(CH₃)₂, —CH₃,—CH₂CH₃, —CH(CH₃)₂, —CF₃, —CH₂CF₃, —OCH₃, —OCH₂CH₃, —OCH(CH₃)₂, —OCF₃,—OCH₂CF₃, —S(O)₂—CH₃, —CONH₂, —CONHCH₃, —NHCONHCH₃, —COCH₃, —COOH, andthe like. In some embodiments, substituted groups are substituted withone, two, or three of the preceding groups. In some embodiments,substituted groups are substituted with one or two of the precedinggroups. In some embodiments, substituted groups are substituted with oneof the preceding groups. Further, unless stated to the contrary, aformula with chemical bonds shown only as solid lines and not as wedgesor dashed lines contemplates each possible isomer, e.g., each enantiomerand diastereomer, and a mixture of isomers, such as racemic or scalemicmixtures.

In some embodiments, a compound of the disclosure is present in acomposition as a salt. In some embodiments, salts are obtained byreacting a compound of the disclosure with acids. In some otherembodiments, pharmaceutically acceptable salts are obtained by reactinga compound of the disclosure with a base. In other embodiments, thecompounds are used as free-acid or free-base form in the manufacture ofthe compositions described herein. The type of salts, include, but arenot limited to: (1) acid addition salts, formed by reacting the freebase form of the compound with a pharmaceutically acceptable: inorganicacid, such as, for example, hydrochloric acid, hydrobromic acid,sulfuric acid, phosphoric acid, metaphosphoric acid, and the like; orwith an organic acid, such as, for example, acetic acid, propionic acid,hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid,lactic acid, malonic acid, succinic acid, malic acid, maleic acid,fumaric acid, trifluoroacetic acid, tartaric acid, citric acid, benzoicacid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid,methanesulfonic acid, ethanesulfonic acid, 1,2-ethanedisulfonic acid,2-hydroxyethanesulfonic acid, benzenesulfonic acid, toluenesulfonicacid, 2-naphthalenesulfonic acid,4-methylbicyclo-[2.2.2]oct-2-ene-1-carboxylic acid, glucoheptonic acid,4,4′-methylenebis-(3-hydroxy-2-ene-1-carboxylic acid), 3-phenylpropionicacid, trimethylacetic acid, tertiary butylacetic acid, lauryl sulfuricacid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylicacid, stearic acid, muconic acid, butyric acid, phenylacetic acid,phenylbutyric acid, valproic acid, and the like; (2) salts formed whenan acidic proton present in the parent compound is replaced by a metalion, e.g., an alkali metal ion (e.g. lithium, sodium, potassium), analkaline earth ion (e.g. magnesium or calcium), or an aluminum ion. Insome cases, the lipid modulating compound described herein are reactedwith an organic base, such as, but not limited to, ethanolamine,diethanolamine, triethanolamine, methylamine, dimethylamine,trimethylamine, ethylamine, diethylamine, triethylamine,N-methylglucamine, dicyclohexylamine, tris(hydroxymethyl)methylamine. Inother cases, the compounds described herein form salts with amino acidssuch as, but not limited to, arginine, lysine, and the like. Acceptableinorganic bases used to form salts with compounds that include an acidicproton, include, but are not limited to, aluminum hydroxide, calciumhydroxide, potassium hydroxide, sodium carbonate, sodium hydroxide, andthe like.

“Addition compound” refers to a complex of two or more completemolecules in which each preserves its fundamental structure and nocovalent bonds are made or broken (for example, hydrates of salts,adducts).

“Aliphatic acid” refers to acids of nonaromatic hydrocarbons. Examplesof aliphatic acids include, but are not limited to, butyric acid,hexanoic acid, propionic acid, octanoic acid, and acetic acid.

“Alkene” refers to an unsaturated linear divalent hydrocarbon moiety ofone to twelve, typically one to six, carbon atoms or a saturatedbranched divalent hydrocarbon moiety of three to twelve, typically threeto six, carbon atoms. Exemplary alkene groups include, but are notlimited to, methylene, ethylene, propylene, butylene, pentylene, and thelike.

“Antagonist” refers to a compound or a composition that attenuates theeffect of an agonist. The antagonist can directly bind reversibly orirreversibly to a region of the receptor in common with an agonist. Anantagonist can also bind at a different site on the receptor or anassociated ion channel. Thus, the term “antagonist” includes afunctional antagonist. A “functional antagonist” refers to a compoundand/or composition that reverses the effects of an agonist by meansother than acting at the same receptor as the agonist, i.e., afunctional antagonist causes a response in the tissue or animal whichopposes the action of an agonist. Examples include agents which haveopposing effects on an intracellular second messenger or on aphysiologic state in an animal (for example, blood pressure).

“Biological activity” as used herein means having an effect on oreliciting or preventing a response from a living cell, tissue, organ orphysiologic activity, such as, but not limited to, altering gene and/orprotein expression, protein phosphorylation, cellular behavior, and/ororgan function.

“Biomarker” as used herein means a measurable indicator of the severityor the presence of a particular disease state. More generally abiomarker is anything that can be used as an indicator of a particulardisease state or some other physiological state of an organism.

“Carboxyl” refers to an organic functional group consisting of a carbonatom double bonded to an oxygen atom and single bonded to a hydroxylgroup.

“Chiral center” (i.e., stereochemical center, stereocenter, orstereogenic center) refers to an asymmetrically substituted atom, e.g.,a carbon atom to which four different groups are attached. The ultimatecriterion of a chiral center, however, is nonsuperimposability of itsmirror image. If an asymmetric center is present in one or moresubstituents, the compound may be in the form of a racemic mixture, asingle enantiomer, a diastereoisomeric mixture, an enantiomericdiastereomer, a meso compound, a pure epimer, or a mixture of epimersthereof.

“Derivative” refers to a compound that is derived from some parentcompound where one atom is replaced with another atom or group of atomsand usually maintains its general structure. For example,trichloromethane (chloroform) is a derivative of methane.

“Dermatitis” as used herein refers to a general or localizedinflammation of the skin, either due to an inherent skin defect, directcontact with an irritating substance, virus, bacteria, animal parasite,fungus or to an allergic reaction. Symptoms of dermatitis include:redness, itching, exudations, pain, fissures, cracks, ulcers, and insome cases blistering of the skin.

“Eczema” as used herein refers to an inflammatory condition of the skincharacterized by redness, itching, and oozing vesicular lesions whichbecome scaly, crusted, or hardened.

“Epithelium (epithelia, plural) or epithelial tissues” as used hereinmeans a type of animal tissue made up of densely packed cells that reston a basement membrane to act as a covering of a free surface such as,but not limited to the surface of the human body; or lining of variousbodily surfaces, such as but not limited to the eyes; or lining variousbody cavities such as but not limited the abdominal cavity; or liningthe lumina of tubular structures within organs, such as but not limitedto the respiratory epithelium, urogenital epithelium.

“Enantiomeric excess” refers to the difference between the amounts ofenantiomers. The percentage of enantiomeric excess (% ee) can becalculated by subtracting the percentage of one enantiomer from thepercentage of the other enantiomer. For example, if the % of(R)-enantiomer is 99% and % of (S)-enantiomer is 1%, the % ee of(R)-isomer is 99%-1% or 98%.

The terms “halo,” “halogen” and “halide” are used interchangeably hereinand refer to fluoro, chloro, bromo, or iodo.

“Haloalkyl” refers to an alkyl group as defined herein in which one ormore hydrogen atom is replaced by the same or different halo atoms. Theterm “haloalkyl” also includes perhalogenated alkyl groups in which allalkyl hydrogen atoms are replaced by halogen atoms. Exemplary haloalkylgroups include, but are not limited to: —CH₂F, —CH₂C₁, —CF₃, —CH₂CF₃,—CH₂CC13, and the like.

“Hetero-substituted alkyl” refers to an alkyl group, as defined herein,that contains one or more heteroatoms such as N, O, or S. Suchheteroatoms can be hydroxy, alkoxy, amino, mono-, di-alkyl amino, thiol,alkylthiol, etc.

“Hydroxyalkyl” refers to an alkyl group, as defined herein, having oneor more hydroxyl sub stituent(s).

“Keto acid” refers to organic compounds that contain a carboxylic acidgroup and a ketone group.

“Leaving group” has the meaning conventionally associated with it insynthetic organic chemistry, i.e., an atom or a group capable of beingdisplaced by a nucleophile and includes halo (such as chloro, bromo, andiodo), alkanesulfonyloxy, arenesulfonyloxy, alkylcarbonyloxy (e.g.,acetoxy), arylcarbonyloxy, mesyloxy, tosyloxy,trifluoromethanesulfonyloxy, aryloxy (e.g., 2,4-dinitrophenoxy),methoxy, N, O-dimethylhydroxylamino, and the like.

“Leukocyte or leukocytes” as used herein refer to a colorless cell thatcirculates in the blood and body fluids and is involved in counteractingforeign substances and infectious disease as well as being causative ofinflammatory diseases. They have also been referred to as white (blood)cells. There are several types, all are amoeboid cells with a nucleus,including lymphocytes, granulocytes, monocytes, and macrophages.

“Ligand” as used herein means a biochemical substance in the form of anucleic acid, protein or peptide that forms a complex with anotherbiomolecule in a cell or tissue to serve a biological purpose.

“Moderate” as used herein means to decrease or increase the quality,quantity, intensity or duration of a biological product or process.

“Peripheral blood mononuclear cell (PBMC)” as used herein means anyperipheral blood cell having a round nucleus. These cells are a subsetof leukocytes and consist of lymphocytes (T cells, B cells, NK cells)and monocytes, whereas erythrocytes and platelets have no nuclei andgranulocytes (neutrophils, basophils, and eosinophils) have multi-lobednuclei.

“Pharmaceutically acceptable excipient” refers to an excipient that isuseful in preparing a pharmaceutical composition that is generally safe,non-toxic, and neither biologically nor otherwise undesirable, andincludes excipients that are acceptable for veterinary use as well ashuman pharmaceutical use.

“Pharmaceutically acceptable salt” of a compound means a salt that ispharmaceutically acceptable and that possesses the desiredpharmacological activity of the parent compound. Such salts include:acid addition salts formed with inorganic acids such as hydrochloricacid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, andthe like; or formed with organic acids such as acetic acid, propionicacid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvicacid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid,fumaric acid, tartaric acid, citric acid, benzoic acid,3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid,methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid,2-hydroxyethanesulfonic acid, benzenesulfonic acid,4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid,4-toluenesulfonic acid, camphorsulfonic acid,4-methylbicyclo[2.2.2]-oct-2-ene-1-carboxylic acid, glucoheptonic acid,3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic acid,lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoicacid, salicylic acid, stearic acid, muconic acid, and the like. A“pharmaceutically acceptable salt” of a compound also includes saltsformed when an acidic proton present in the parent compound is eitherreplaced by a metal ion, e.g., an alkali metal ion, an alkaline earthion, or an aluminum ion; or coordinates with an organic base such asethanolamine, diethanolamine, triethanolamine, tromethamine,N-methylglucamine, and the like.

Pharmaceutically acceptable vehicle means a carrier or inert medium usedas a solvent (or diluent) in which the medicinally active agent isformulated and or administered.

The terms “pro-drug” and “prodrug” are used interchangeably herein andrefer to any compound which releases an active parent drug according toFormula 1, or a pharmaceutically acceptable salt or solvate of Formula1, in vivo when such prodrug is administered to a mammalian subject.Prodrugs of a compound of Formula 1 are prepared by modifying one ormore functional group(s) present in the compound of Formula 1 in such away that the modification(s) may be cleaved in vivo to release theparent compound. Prodrugs include compounds of Formula 1 wherein ahydroxy group in a compound of Formula 1 is bonded to any group that maybe cleaved in vivo to regenerate the free hydroxyl, aliphatic alcohol,amino, or sulfhydryl group, respectively. Examples of prodrugs include,but are not limited to, esters (e.g., acetate, formate, glycol andbenzoate derivatives of Formula 1) and the like. For example, thecompound according to Formula 1 that is methyl3-(4-hydroxyphenoxy)-3-methyl-butanoate can be reacted under acidicconditions with 2-hydroxybenzoic acid to produce,[4-(3-methoxy-1,-1-dimethyl-3oxo-propoxy)2-hydoxybenzoate an esterprodrug that will be hydrolyzed to 2-hydroxybenzoic acid and thestarting compound by esterase enzymes in tissues. The transformationfrom prodrug to a compound of Formula 1, or a pharmaceuticallyacceptable salt or solvate thereof, may occur by various mechanisms,such as via hydrolysis in blood. A prodrug of the compound of Formula 1may be formed in a conventional manner according to methods known in theart. A thorough discussion of prodrugs is provided by V. Stella inPro-drugs as Novel Delivery Systems, Vol. 14 of the A.C.S. SymposiumSeries (Stella (1975)), and in Bioreversible Carriers in Drug Design(Roche (1987)), both of which are incorporated herein by reference.

“Pro-inflammatory cytokine” refers to a type of cytokine (i.e. a proteinsignaling molecule) that is secreted from leukocytes, monocytes andother non-leukocyte cell types; such as but not limited epithelialcells, that promote inflammation by their biological effect on othercells and tissue in mammalian organisms. Non-limiting examples ofpro-inflammatory cytokines are: Interleukin 1 (IL-1; IL-la & IL-1b),Interleukin 6 (IL-6), Interleukin 13 (IL-13), Tumor Necrosis Factoralpha (TNF-alpha), Interferon gamma (IFN-gamma), and Interleukin 8(IL-8).

The term “prophylaxis” of a state, disorder, disease or condition asused herein refers to prevention of the appearance of clinical symptomsof the state, disorder, disease or condition developing in a patientthat is predisposed to the state, disorder, disease, or condition.

“Protecting group” refers to a moiety, with the exception of alkylgroups, that when attached to a reactive group in a molecule masks,reduces, or prevents that reactivity. Examples of protecting groups canbe found in T. W. Greene and P. G. M. Wuts, Protective Groups in OrganicSynthesis, 3rd edition, John Wiley & Sons, New York, 1999, and Harrisonand Harrison et al., Compendium of Synthetic Organic Methods, Vols. 1-8(John Wiley and Sons, 1971-1996), which are incorporated by referenceherein in their entirety. Representative hydroxy protecting groupsinclude acyl groups, benzyl and trityl ethers, tetrahydropyranyl ethers,trialkylsilyl ethers, and allyl ethers. Representative amino protectinggroups include, formyl, acetyl, trifluoroacetyl, benzyl,benzyloxycarbonyl (CBZ), tert-butoxycarbonyl (Boc), trimethyl silyl(TMS), 2-trimethylsilyl-ethanesulfonyl (SES), trityl and substitutedtrityl groups, allyloxycarbonyl, 9-fluorenylmethyloxycarbonyl (FMOC),nitro-veratryloxycarbonyl (NVOC), and the like.

“Corresponding protecting group” means an appropriate protecting groupcorresponding to the heteroatom (i.e., N, O, P, or S) to which it isattached.

“Signal transduction” or “signaling pathway activity” refers to abiochemical causal relationship generally initiated by a protein-proteininteraction such as binding of a biological active factor to a receptor,resulting in transmission of a signal from one portion of a cell toanother portion of a cell. In general, the transmission can involvespecific phosphorylation of one or more tyrosine, serine, or threonineresidues on one or more protein components such as enzymes ortranscription factors (i.e. intracellular secondary messengers) in theseries of reactions causing signal transduction (often referred to as acascade) that results in measurable changes to the cell. Penultimatecellular processes typically include nuclear events, resulting in achange in gene expression. Terminal events of signal transductioncascade result in changes in cellular activity such as but not limitedto, alterations in protein products produced and/or secreted by thecell, changes in cellular behavior characteristics of division,motility, adherence, etc.

“Stereoisomer” means molecules that have the same molecular formula,molecular weight and sequence of bonded atoms (constitution), but differin the three-dimensional orientations of their atoms in space. Bydefinition, molecules that are stereoisomers of each other represent thesame structural isomer. The chemical definitions and conventions usedherein generally follow S. P. Parker, Ed., McGraw-Hill Dictionary ofChemical Terms (1984) McGraw-Hill Book Company, New York; and Eliel, E.and Wilen, S., “Stereochemistry of Organic Compounds”, John Wiley &Sons, Inc., New York, 1994.

“A therapeutically effective amount” means the amount of a compoundthat, when administered to an individual for treating a disease, issufficient to effect such treatment for the disease, as defined below.The “therapeutically effective amount” will vary depending on thecompound, the disease and its severity or affected organ or tissue andthe age, weight, etc., of the individual to be treated.

“Tautomer” or “tautomeric form” means structural isomers of differentenergies which are interconvertible via a low energy barrier. Forexample, proton tautomers (also known as prototropic tautomers) includeinterconversions via migration of a proton, such as keto-enol andimine-enamine isomerizations. Valence tautomers include interconversionsby reorganization of some of the bonding electrons. The compounds of thepresent invention according to Formula 1 can exist in differenttautomeric states depending on the environment of the particularcompound, such as the acidity or alkalinity (i.e. pH) of the solution inwhich they are dissolved.

“Topical or topically applied” as used herein means the direct deliveryor application of the active drug ingredient such as compounds accordingto Formula 1 directly to the surface of the epithelial tissue by meansof a spray, cream, ointment, shampoo, lotion, solution or other suitabledelivery solvent or vehicle. Various methods of topical drug deliverysuch as; but not limited to mechanical application, instillation,inhalation, patches, and the like are known to those skilled in the art,are commonly used for topical application, and are implicit in theconcept of topical application as used herein.

“Treating” or “treatment” of a disease means inhibiting the disease,i.e., arresting or reducing the pathophysiologic process or processes ofthe disease or its clinical symptoms; or relieving the disease, i.e.,causing regression of the pathophysiologic process or processes ofdisease or reducing the clinical manifestations of the pathophysiologicprocess or processes of the specific disease.

In some embodiments, a compound of the disclosure is present in acomposition as a salt. In some embodiments, salts are obtained byreacting a compound of the disclosure with acids. In some otherembodiments, pharmaceutically acceptable salts are obtained by reactinga compound of the disclosure with a base. In other embodiments, thecompounds are used as free-acid or free-base form in the manufacture ofthe compositions described herein. The type of salts, include, but arenot limited to: (1) acid addition salts, formed by reacting the freebase form of the compound with a pharmaceutically acceptable: inorganicacid, such as, for example, hydrochloric acid, hydrobromic acid,sulfuric acid, phosphoric acid, metaphosphoric acid, and the like; orwith an organic acid, such as, for example, acetic acid, propionic acid,hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid,lactic acid, malonic acid, succinic acid, malic acid, maleic acid,fumaric acid, trifluoroacetic acid, tartaric acid, citric acid, benzoicacid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid,methanesulfonic acid, ethanesulfonic acid, 1,2-ethanedisulfonic acid,2-hydroxyethanesulfonic acid, benzenesulfonic acid, toluenesulfonicacid, 2-naphthalenesulfonic acid,4-methylbicyclo-[2.2.2]oct-2-ene-1-carboxylic acid, glucoheptonic acid,4,4′-methylenebis-(3-hydroxy-2-ene-1-carboxylic acid), 3-phenylpropionicacid, trimethylacetic acid, tertiary butylacetic acid, lauryl sulfuricacid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylicacid, stearic acid, muconic acid, butyric acid, phenylacetic acid,phenylbutyric acid, valproic acid, and the like; (2) salts formed whenan acidic proton present in the parent compound is replaced by a metalion, e.g., an alkali metal ion (e.g. lithium, sodium, potassium), analkaline earth ion (e.g. magnesium, or calcium), or an aluminum ion. Insome cases, the lipid modulating compound described herein are reactedwith an organic base, such as, but not limited to, ethanolamine,diethanolamine, triethanolamine, methylamine, dimethylamine,trimethylamine, ethylamine, diethylamine, triethylamine,N-methylglucamine, dicyclohexylamine, tris(hydroxymethyl)methylamine. Inother cases, the compounds described herein form salts with amino acidssuch as, but not limited to, arginine, lysine, and the like. Acceptableinorganic bases used to form salts with compounds that include an acidicproton, include, but are not limited to, aluminum hydroxide, calciumhydroxide, potassium hydroxide, sodium carbonate, sodium hydroxide, andthe like.

In the scope of the embodiments, the compounds described herein includefurther forms of the compounds such as pharmaceutically acceptablesalts, solvates (including hydrates), amorphous phases, partiallycrystalline and crystalline forms (including all polymorphs), prodrugs,metabolites, N-oxides, isotopically-labeled, epimers, pure epimers,epimer mixtures, enantiomers including, but not limited to, singleenantiomers and enantiomeric diastereomers, meso compounds,stereoisomers, racemic mixtures, and diastereoisomeric mixtures.Compounds described herein having one or more double bonds includecis/trans isomers, E/Z isomers and geometric isomers.

In some embodiments, sites on the compounds disclosed herein aresusceptible to various metabolic reactions. Therefore, incorporation ofappropriate substituents at the places of metabolic reactions willreduce, minimize or eliminate the metabolic pathways. In specificembodiments, the appropriate substituent to decrease or eliminate thesusceptibility of the aromatic ring to metabolic reactions is, by way ofexample only, a halogen, deuterium, or an alkyl group. Examples of suchsubstituents can be found in Burger's Medicinal Chemistry, DrugDiscovery and Development, 8 Volume Set (Abraham (2010)) and in Foye'sPrinciples of Medicinal Chemistry (Lemke (2012)).

In some embodiments, sites on the compounds disclosed herein are notsusceptible to various metabolic reactions. Therefore, incorporation ofappropriate substituents at or near or distant from the places of a lackof metabolic reactions will modulate, enhance, or maximize the metabolicpathways. In specific embodiments, the appropriate substituent(metabolic handle) to enhance, or maximize the susceptibility of thearomatic ring to metabolic reactions is, by way of example only, is aphenolic or methoxy or carboxylate group. Examples of such substituentscan be found in Burger's Medicinal Chemistry, Drug Discovery andDevelopment, 8 Volume Set (Abraham (2010)) and in Foye's Principles ofMedicinal Chemistry (Lemke (2012)).

Throughout the specification, groups and substituents thereof can bechosen by one skilled in the field to provide stable moieties andcompounds.

I. METHODS OF SYNTHESIS

The compounds of Formula 1 may be prepared by the methods describedbelow, together with synthetic methods known in the art of organicchemistry, or modifications and transformations that are familiar tothose of ordinary skill in the art. Various starting materials,intermediates, and reagents may be purchased from commercial sources ormade according to literature methods or adaptations thereof. Althoughother reagents, compounds or methods can be used in practice or testing,generalized methods for the preparation of the compound of Formula 1 areillustrated by the following descriptions and reaction Schemes. Themethods disclosed herein, including those outlined in the Schemes,descriptions, and Examples are for intended for illustrative purposesand are not to be construed in any manner as limitations thereon.Various changes and modifications will be obvious to those of skill inthe art given the benefit of the present disclosure and are deemed to bewithin the spirit and scope of the present disclosure as further definedin the appended claims.

Although specific embodiments of various aspects of the invention willbe described with reference to the Schemes, Preparations, and/orExamples, it should be understood that such embodiments are by way ofexample only and are merely illustrative of a small number of the manypossible specific embodiments which can represent applications of theprinciples of the present disclosure. The starting materials used forthe synthesis of compounds described herein can be obtained fromcommercial sources, such as Aldrich Chemical Co. (Milwaukee, Wis.),Sigma Chemical Co. (St. Louis, Mo.), or the starting materials can besynthesized. The compounds described herein and other related compoundshaving different substituents can be synthesized using techniques andmaterials known to those of skill in the art, such as described, forexample, in March's Advanced Organic Chemistry: Reactions, Mechanisms,and Formula (Smith (2013)), Design and Strategy in Organic Synthesis(Hanessian (2013)) Greene's Protective Groups in Organic Synthesis (Wuts(2006)) and Fiesers' Reagents for Organic Synthesis (Volumes 1-27) (Ho(2013)), each of which are incorporated by reference in their entirety.

General methods for the preparation of the compounds as disclosed hereinmay be derived from known reactions in the field and the reactions maybe modified by the use of appropriate reagents and conditions, as wouldbe recognized by the skilled person, for the introduction of the variousmoieties found in the formulae as provided herein.

[87] The intermediate products described can be recovered by extraction,evaporation, or other techniques known in the art. The crude materialsmay then be optionally purified by chromatography, HPLC,recrystallization, trituration, distillation, or other techniques knownin the art. In the discussions below, the following abbreviations wereused: THF (tetrahydrofuran), DMF (N,N-dimethylformamide), BOC(tert-butoxycarbonyl), Cbz (carbobenzoxy), Cs₂CO₃ (Cesium Carbonate),DEPC (diethylcyano-phosphate), LDA (lithium Diisopropylamide), NMP(N-Methyl-2-pyrrolidone), TEA or NEt₃ (triethyl amine), p-TsOH(p-toluene sulfonic acid), i-PrOAc (isopropyl acetate), HOB T(1-hydroxybenzo-triazole), EDC (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide), and EtOH (ethanol).

As would be appreciated by those skilled in the art, some of the methodsuseful for the preparation of such compounds, as discussed above, mayrequire protection of a particular functionality, e.g., to preventinterference by such functionality in reactions at other sites withinthe molecule or to preserve the integrity of such functionality. Theneed for, and type of, such protection is readily determined by oneskilled in the art and will vary depending on, for example, the natureof the functionality and the conditions of the selected preparationmethod. Methods of introducing and removing protecting groups are wellknown to those of ordinary skill in the art and are described inGreene's Protective Groups in Organic Synthesis (Wuts (2006)). Alternatereagents, starting materials, as well as methods for optimizing oradapting the procedures described herein would also be readilydetermined by one skilled in the art.

In Scheme 1 a variety of means are used to generate a phenolate from acommercially available phenol which then displaces the bromine of theR₃R₄ intermediate to give the product shown. Other phenols are easilyavailable by synthesis to those skilled in the art. Other atoms to bedisplaced from the R₃R₄ component are also envisaged including chlorine,iodine and the so-called Mitsonobu adduct derived from treatment of thecorresponding alcohol with DEAD (diazoethyldicarboxylate).

Scheme 2 outlines a similar set of transformations. Again, a variety ofmeans are used to generate a phenolate from a commercially availablephenol which then displaces the bromine of the R₃R₄ intermediate to givethe product shown. Other phenols are easily available by synthesis tothose skilled in the art. Other atoms to be displaced from the R₃R₄component are also envisaged including chlorine, iodine and theso-called Mitsonobu adduct derived from treatment of the correspondingalcohol with DEAD.

Scheme 3 outlines the general synthesis of representative compounds.Again, a variety of means are used to generate a phenolate from acommercially available phenol which then displaces the bromine of theR₃R₄ intermediate to give the product shown. Other phenols are easilyavailable by synthesis to those skilled in the art. Other atoms to bedisplaced from the R₃R₄ component are also envisaged including chlorine,iodine and the so-called Mitsonobu adduct derived from treatment of thecorresponding alcohol with DEAD.

The transformations shown in either Scheme 1 or Scheme 2 or Scheme 3 canbe carried out with racemic material or material which has partialenrichment of one enantiomer over the other as well as optically purematerials.

Scheme 4 is an alternative synthesis which is only suitable for thelimited case where n and p=0 in the generic structure, Formula 1, and isnot suitable for the generation of enantiomers at the possibleasymmetric center adjacent to R₃ and R₄. In this case the phenolategenerated reacts with a ketone carrying R₃ and R₄ followed by reactionof the intermediate with the anion derived from chloroform andhydrolysis to give the desired carboxylic acid. Acid catalyzedesterification then gives the esters shown.

Scheme 5, Scheme 6, and Scheme 7 illustrate how the Suzuki-Miyaurareaction (Molander et al. (2014)) can be used to handle some of the morechallenging substituents envisaged for the central aryl ring withoutgoing through the phenol as the penultimate/final intermediate. Theoptimally substituted aryl boronate and the suitably substituted arylbromide participate in a CO trapping reaction assisted by Pd(acac)₂.

Again, these transformations can be carried out with racemic materials,partially optically enriched materials and optically pure materials.

Preparation of the example compounds were prepared as set forth aboveand/or with standard procedures well known to those skilled in the art.Example compounds are set forth in Tables 1-13.

TABLE 1 Structure

TABLE 2 Structure

TABLE 3 Structure

TABLE 4 Structure

TABLE 5 Structure

TABLE 6 Structure

TABLE 7 Structure

TABLE 8 Structure Structure Structure Structure Structure

369 370 371 372 373

374 375 376 377 378

379 380 381 382 383

384 385 386 387 388

389 390 391 392 393

394 395 396 397 398

399 400 401 402 403

404 405 406 407 408

409 410 411 412 413

414 415 416

TABLE 9 Structure Structure Structure Structure Structure

417 418 419 420 421

422 423 424 425 426

427 428 429 430 431

432 433 434 435 436

437 438 439 440 441

442 443 444 445 446

447 448 449 450 451

452 453 454 455 456

457 458 459 460 461

472 463 464

TABLE 10 Structure Structure Structure Structure Structure

465 466 467 468 469

470 471 472 473 474

475 476 477 478 479

480 481 482 483 484

485 486 487 488 489

490 491 492 493 494

495 496 497 498 499

500 501 502 503 504

505 506 507 508 509

510 511 512 513 514

515 516 517 518 519

520 521 522 523 524

TABLE 11 Structure Structure Structure Structure Structure

525 526 527 528 529

530 531 532 533 534

535 536 537 538 539

540 541 542 543 544

545 546 547 548 549

550 551 552 553 554

555 556 557 558 559

560 561 562 563 564

565 566 567 568 569

570 571 572

TABLE 12 Structure Structure Structure Structure Structure

573 574 575 576 577

578 579 580 581 582

583 584 585 586 587

588 589 590 591 592

593 594 595 596 597

598 599 600 601 602

603 604 605 606 607

608 609 610 611 612

613 614 615 616 617

618 619 620

TABLE 13 Structure Structure Structure Structure Structure

621 622 623 624 625

626 627 628 629 630

631 632 633 634 635

636 637 638 639 640

641 642

The formulations described in this application are useful fordermatological and cosmetic conditions and may be formulated as apharmaceutical composition and administered to a mammal, such as a humanpatient, in a variety of forms adapted to a chosen route ofadministration, i.e. topically, intralesionally, or subcutaneously. Theformulations described in this application useful for endothelial tissueconditions may also be formulated as a pharmaceutical composition andadministered to a mammal, such as a human patient, intravenously. Itshould be understood that the invention is not limited by the chosenroute of administration. The compound present may be administered aloneor in combination with one or more other therapeutic agents.

In a typical embodiment the compound is administered as a formulation inassociation with a pharmaceutically acceptable carrier. The choice ofcarrier largely depends on factors such as the particular mode ofadministration, the effect of the carrier on solubility and stability,and the nature of the dosage form.

In some embodiments, the compound will be formulated with a carriersuitable for administration directly to the skin or hair.

In yet another embodiment, the compound is formulated with a carriersuitable for intravenous administration.

In other embodiments, the compound is topically applied to a subject.Topical application is especially appropriate for the treatment of acne,rosacea, excess sebum, oily skin or hair, and shiny or greasy lookingskin. In certain embodiments, topical application refers to applicationof a compound, and optional carrier, directly to the skin and/or hair.The topical composition according to the present invention can be in theform of solutions, lotions, salves, creams, ointments, liposomes,sprays, gels, foams, roller sticks, or any other formulation routinelyused in dermatology.

In other embodiments, compositions of the invention may be solid orsemi-solid formulations which are suitable for use as cleansing soaps,gels, or bars. These compositions are prepared according to the usualmethods and may optionally contain additional excipients such asmoisturizers, colorants, fragrances, and the like.

The compound may also be formulated for application to the hair in theform of aqueous, alcoholic or aqueous-alcoholic solutions, or in theform of creams, gels, emulsions or mousses, or alternatively in the formof aerosol compositions also comprising a propellant under pressure. Thecomposition according to the invention can also be a hair carecomposition, and in particular a shampoo, a hair-setting lotion, atreating lotion, a styling cream or gel, a dye composition, a lotion orgel for preventing hair loss, etc. The amounts of the excipients in thevarious compositions according to the invention are those conventionallyused in the fields considered.

Pharmaceutical compositions suitable for the delivery of compounds ofthe present invention and methods for their preparation will be readilyapparent to those skilled in the art. Such compositions and methods fortheir preparation may be found, for example, in Remington'sPharmaceutical Sciences, 19th Edition (Gennaro (1995)).

II. METHODS OF INHIBITION OF CCL5 PRODUCTION BY CELLS

The compounds and compositions described herein inhibit the productionof CCL5 by various types of mammalian cells present in epithelial andendothelial tissues. Non-limiting examples are inhibition of productionof CCL5 by keratinocytes, respiratory epithelial cells, fibroblasts, andendothelial cells under conditions known to increase CCL5 geneexpression and subsequent release CCL5 into the cellular or tissuemilieu, as measured by various assays including immune based proteinquantitation.

III. METHODS OF TREATMENT

CCL5 acts as a cytokine when secreted by cells and participates inpro-inflammation response, angiogenesis, leukocyte adhesion, andextravasation into tissues. Increased blood and/or tissue levels of CCL5have been shown to contribute to disease pathogenesis and progression.Increased tissue or blood levels of CCL5 are directly contributory tothe pathogenesis of: autoimmune diseases; inflammatory diseases;auto-inflammatory conditions; pain conditions; respiratory ailments;airway and pulmonary conditions; gastrointestinal disorders; allergicdiseases; atopic disorders; eczematous conditions; infection-baseddiseases; trauma and tissue injury-based conditions; fibrotic diseases;ophthalmic/ocular diseases; joint, muscle, and bone disorders;skin/dermatological diseases; renal diseases; genetic diseases;hematopoietic diseases; liver diseases; oral diseases; metabolicdiseases, including diabetes (e.g. Type II) and complications thereof;proliferative diseases; cardiovascular conditions; vascular conditionsincluding restenosis; neuro-inflammatory conditions; neurodegenerativeconditions; cancer; and pulmonary conditions. Thus, inhibition of CCL5production has implications for treatment of many disorders with diverseprimary etiologies.

With regard to the increased levels of secreted pro-inflammatorycytokines and chemokines produced by leukocytes, CCL5 through itsinteraction with its receptor(s) (known as CCL5, CCL3, CCL3L1, andCCL4), present on many types of leukocytes, causes the increasedproduction of cytokines produced by leukocytes and especially monocytessuch as Th1 and Th2 cells. Among the pro-inflammatory cytokines inducedby CCL5 are: IL-1 beta, TNF-alpha, IL-6, and IFN-gamma. These increasedpro-inflammatory cytokines and chemokines act to induce pathogenicprocesses in various non-leukocyte cells and are responsible for acuteand chronic diseases and their complications such as, but not limitedto; Atopic disorders, Autoimmune diseases, Carcinoma, Cardiac disorders,Dermatologic diseases, Fibrosis, Gastrointestinal disorders, Hepaticdiseases, Infectious diseases, Inflammatory disorders, Metabolicdisorders (e.g. diabetes), Nephropathies, Neoplasia, Neurodegenerativedisorders, Ophthalmologic disorders, Osteoporosis, Pulmonary diseases,Urinary tract disorders, Vascular conditions including restenosis, andothers. Inhibitions of the production of CCL5 by the compounds of thepresent invention reduce the tissue levels of these pro-inflammatorycytokines and chemokines produced by leukocytes and other cell types.

As used in this application, the terms “co-administered” or“co-administration” refer to a dosing regimen where the compound ofFormula 1 is administered with a second therapeutic agent, typicallyhaving a differing mechanism of action, to promote a desired result. Itshould be understood that “co administration” is not limited by theroute(s) of administration and can refer to simultaneous dosing, dosingat different times during a single day, or even dosing on differentdays. The compounds can be administered separately or can be combinedinto a single formulation (i.e. fixed combination).

In an embodiment of the present invention the compounds according toFormula 1 may be used either simultaneously or sequentially incombination with a second compound such as, but not limited to, thoselisted below to reduce the production of CCL5 by certain cells such as,but not limited to: keratinocytes, fibroblasts, respiratory epithelialcells, and endothelial cells under conditions that stimulate CCL5production by such cells.

Non-steroidal anti-inflammatory drugs, such as but not limited to:aspirin, choline salicylate, celecoxib, acetaminophen, diclofenac,flurbiprofen, ibuprofen, indomethacin, ketoprofen, meclofenamate,mefenamic acid, nabumetone, naproxen, piroxicam, rofecoxib, salicylates,sulindac, tolmetin, and valdecoxib.

Immunomodulatory agents, such as but not limited to: methotrexate,azathioprine, mitoxantrone, cladribin, cyclophosphamide, tacrorimus,methotrexate, cyclosporine, and hydroxychloroquine.

Antimalarials, such as but not limited to: chloroquine, quinine,amodiaquine, pyrimethamine, proguanil, mefloquine, atovaquone,primaquine, artemisinin, and halofantrine.

Antibiotics, such as but not limited to: sulfonamides, clindamycin,members of the tetracycline family (including minocycline anddoxycycline), erythromycin, and dapsone.

Anti-TNF alpha agents, such as but not limited to: infliximab,adalimumab, certolizumab pegol, golimumab, thalidomide, lenalidomide,pomalidomide, and etanercept.

Anti-CD20 agents, such as but not limited to: rituximab, obinutuzumab,Ibritumomab tiuxetan, and tositumomab.

Antidiarrheals, such as but not limited to: lidamidine, diphenoxylate,loperamide, and quercetin.

Antidepressants, such as but not limited to: amitriptyline,clomipramine, doxepin nortriptyline, and trimipramine.

Antipsychotics, such as but not limited to: droperidol, pimozide,chlorpromazine, thiothixene, loxapine, molindone, quetiapine,risperidone, sertindole, and zotepine.

Antifungals, such as but not limited to: clotimazole, flucisoconazole,abafungin, micafugin, terbinafine, ciclopirox, and tolnaftate.

Antihelminthics, such as but not limited to: mebendazole, levamisole,abamectin, and suramine

T lymphocyte activation inhibitors, such as but not limited to:voclosporin, peroxynitrite, and dasatinib.

Anti-IL-1 agents, such as but not limited to: anakinra and IL-1Ra.

Glucocorticoids, such as but not limited to: methyl prednisolone,prednisolone, dexamethasone, betamethasone, fluticasone propionate,budesonide, flunisolide, mometasone furoate, triamcinolone acetonide,rofleponide, ciclesonide, and butixocort propionate.

Anti-cytokine/chemokine monoclonal antibodies, such as but not limitedto: basiliximab, daclizumab, and secukinumab.

Sex steroids and receptor modulators, such as but not limited to:progesterone, progestins, androgen, estrogen, mifepristone, andmisoprostil.

Anti-cellular surface receptor monoclonal antibodies directed againstcell surface receptors such as but not limited: CCR1, CCR3, CCL3L1,CCL4, CCR5, IL7Ra, and TSLPR.

Aminosalicylic acid derivatives such as but not limited to:sulfasalazine and mesalazine.

Anticholinergic agents, such as but not limited to: ipratropium,oxitropium, tiotropium, dextromethorphan, revatropate, pirenzepine,darifenacin, oxybutynin, mecamylamine, terodiline, tolterodine,otilonium, trospium chloride, and solifenacin.

Adrenergic agonists, such as but not limited to: salmeterol, salbutamol,clonidine, oxymetazoline, and dolbutamine.

Cholineric agonists, such as but not limited to: carbachol, epibatidine,galantamine, nicotine, and varenicline,

Corticosteroids, such as but not limited to: cortisone andhydrocortisone.

Antineoplastic chemotherapeutic agents, such as but not limited to:cisplatin cyclophosphamide, bleomycin, doxorubicin, etoposide, folinicacid, and vincristine.

Phosphodiesterase inhibitors, such as but not limited to: mesembrenone,rolipram, Ibudilast, piclamilast, luteolin, drotaverine, roflumilast,cilomilast, apremilast, and crisaborole.

Leukotriene pathway modulators, such as but not limited to:3-[3-butylsulfanyl-1-[(4-chlorophenyl)methyl]-5-propan-2-yl-indol-2-yl]-2,2-dimethyl-propanoicacid, baicalein, caffeic acid, curcumin, hyperforin, and zileuton.

Monoclonal antibodies directed against human immunoglobulins, such asbut not limited to: omalizumab.

Adrenergic antagonists, such as but not limited to: alfluosin, idazoxan,labetalol, phentolamine, trazadone, propranolol, and atenolol.

Calcium channel antagonists, such as but not limited to: amelodipine,nifedapine, verapamil, diltiazem, and mibefradil.

Dopamine agonists, such as but not limited to: aripiprazole,bromocriptine, bupropion, cabergoline, lisuride, and roxindole.

Serotonin agonists, such as but not limited to: cabergoline, cisapride,gepirone, lorcaserin, and naratriptan.

Dopamine antagonists, such as but not limited to: amoxipine, bromopride,butaclamol, eticlopride, olanzapine, tiapride, and ziprasidone.

Serotonin antagonists, such as but not limited to: cyproheptadine,ketanserin, metergoline, methdilazine, oxetorone, and tropisetron.

Monoamine reuptake inhibitors, such as but limited to: amineptine,citalopram, edivoxetine, hyperforin, mazindol, and viloxazine.

Protease inhibitors, such as but not limited to: amastatin, bestatin,and gabexate.

Histamine receptor antagonists, such as but not limited to: acrivastine,brompheniramine, cetirizine, cimetidine, ciproxifan, clobenprobit,cyclizine, carebastine, cyproheptadine, ebastine, epinastine,efletirizine, fexofenadine, and thioperamide.

Proton pump inhibitors, such as but not limited to; omeprazole,lansoprazole, pantoprazole, and rabeprazole.

HMG-CoA reductase inhibitors, such as but not limited to: atorvastatin,fluastatin, lovastatin, and simvastatin.

Retinoids such as, but not limited to, etretinate, tretinoin, retinol,retinyl palmitate, adapalene, tazarotene, and aliretinoin.

Administration of the therapeutic agent may be by any suitable means. Insome embodiments, the one or more therapeutic agents are administered byoral administration. In some embodiments, the one or more therapeuticagents are administered by transdermal administration. In someembodiments, the one or more therapeutic agents are administered byinjection or intravenous infusion. In one embodiment, the one or moretherapeutic agents are administered topically to a mucosal, dermal, orocular tissue.

If combinations of agents are administered as separate compositions,they may be administered by the same route or by different routes. Ifcombinations of agents are administered in a single composition, theymay be administered by any suitable route. In some embodiments,combinations of agents are administered as a single composition by oraladministration. In some embodiments, combinations of agents areadministered as a single composition by transdermal administration. Insome embodiments, the combinations of agent are administered as a singlecomposition by injection. In some embodiments, the combinations of agentare administered as a single composition topically.

In one embodiment of the present invention the compounds of Formula 1may contain asymmetric or chiral centers and, therefore, exist indifferent stereoisomeric forms. For example, 2,2,2-trifluoroethyl2-(4-methylphenoxy)propanoate is a compound according Formula 1 thatpossesses a chiral center at carbon atom number 8 and thus has twostereoisomer forms. It is intended that all stereoisomeric forms of thecompounds of Formula 1 form part of the present invention, including butnot limited to: diastereomers, enantiomers, and atropisomers as well asmixtures thereof, such as racemic mixtures. In addition, the presentinvention embraces all geometric and positional isomers. For example, ifa compound of Formula 1 incorporates a double bond or a fused ring, boththe cis- and trans-forms, as well as mixtures, are embraced within thescope of the invention. Both the single positional isomers and mixtureof positional isomers are also within the scope of the presentinvention.

In one embodiment of the present invention, compounds of Formula 1 mayexist in different tautomeric forms, and all such forms are embracedwithin the scope of the invention, as defined by the claims.

The dose and dosing regimens of the compound present in the inventionmay be adjusted to provide the optimum desired response in accordancewith methods and practices well known in the therapeutic arts. Forexample, a single bolus dose may be administered, or several divideddoses may be administered over time. The dose may also be proportionallyreduced or increased as indicated by the exigencies of the therapeuticsituation. The appropriate dosing regimen, the amount of each doseadministered and/or the intervals between doses will depend upon anumber of factors, including: the compound, the type of pharmaceuticalcomposition, the characteristics of the subject in need of treatment andthe severity of the condition being treated.

The dose of the compound will vary, but as a general guideline fordermatological administration, the compound will be present in adermatologically acceptable formulation in a therapeutically effectivedose in an amount of from about 0.001 mg/kg to about 1000 mg/kg/bodyweight per day of a compound provided herein. The pharmaceuticalcompositions therefore should provide a dosage of from about 0.001mg/kg/body weight to about 1000 mg/kg/body weight of the compound forsome conditions. In another embodiment of the present invention thepharmaceutical dosage unit forms are prepared to provide a preparationfor topical application containing 0.01 to 50 w/w %, and more typicallyfrom about 0.1 to 10 w/w %. In yet other embodiments the pharmaceuticaldosage unit forms are prepared to provide a preparation for topicalapplication containing from 0.01% to 30% (w/v) of the compounds.

In some embodiments, the formulation may be applied to the affected areafrom 1 to 4 times daily. A “dermatologically acceptable formulation” isone that may be applied to the skin or hair and will allow the drug todiffuse to the site of action.

The dose of the compound will also vary for endothelial tissueadministration, but as a general guideline for endothelialadministration, the compound will be present in an endotheliallyacceptable formulation in a therapeutically effective dose in an amountof from about 500 picomolar (pM) to 300 millimolar (mM).

The skilled artisan can also be expected to readily determine themaximum tolerable dose, the therapeutically effective amount whichprovides a detectable therapeutic benefit to a patient, and the temporalrequirements for administering each agent to provide a detectabletherapeutic benefit to the patient. Accordingly, while certain dose andadministration regimens are exemplified herein, these examples in no waylimit the dose and administration regimen that may be provided to apatient in practicing the present invention.

The determination of optimal dosages for a particular patient iswell-known to those skilled in the art. Certain non-limiting examples ofpharmaceutically acceptable vehicles suitable for topical administrationinclude propylene glycol:transcutanol:ethanol (20:20:60, v/v/v) andpropylene glycol:ethanol (30:70, v/v). In some embodiments, the compoundof Formula 1 may be present at concentrations of between about 1.5% toabout 2.0% (w/v).

In another embodiment, the medicinal and cosmetic formulationscontaining the compound and any additional therapeutic agents willtypically be packaged for retail distribution (i.e. an article ofmanufacture or a kit). Such articles will be labeled and packaged in amanner to instruct the patient how to use the product. Such instructionswill include the condition to be treated, duration of treatment, dosingschedule, etc. The compound(s) of Formula 1 may also be admixed with anyinert carrier and utilized in laboratory assays in order determine theconcentration of the compounds within the serum, urine, etc., of thepatient as is known in the art. The compound may also be used as aresearch tool.

While the invention has been described in connection with specificembodiments thereof, it will be understood that it is capable of furthermodifications and this application is intended to cover any variations,uses, or adaptations of the invention following, in general, theprinciples of the invention and including such departures from thepresent disclosure as come within known or customary practice within theart to which the invention belongs. The following examples andbiological data are being presented in order to further illustrate theinvention. This disclosure should not be construed as limiting theinvention in any manner.

For all of the foregoing embodiments, each embodiment disclosed hereinis contemplated as being applicable to each of the other disclosedembodiments. Those skilled in the art will readily appreciate that thespecific Experimental Details which follow are only illustrative of theinvention as described more fully in the claims which follow thereafter.

IV. EXAMPLES

To determine the ability of compounds according to Formula 1 to inhibitCCL5 production using a functional assay, cultivated humankeratinocytes, endothelial cells were treated with known stimulants ofCCL5 production and the gene expression and production of CCL5 proteinwere determined by well-known methods as described below.

Human keratinocyte cells (NEHK) from healthy volunteers were obtainedand cultivated at 1×10⁶ cells/ml using 6 well plates in NEHK-GM mediaaccording to supplier's protocol (MatTek Inc., Ashland, Mass. 01721).Human bronchial epithelial cells (NHBE) were obtained and cultivated at1×10⁶ cells/ml using 6 well plates in NHBE-GM media according tosupplier's protocol (MatTek Inc. Ashland, Mass. 01721). Human umbilicalvein endothelial cells (HUVEC) were obtained and cultivated at 1×10⁶cells/ml using 6 well plates in supplemented Medium 200 according tosupplier's protocol (ThermoFisher-Invitrogen Carlsbad Calif. 92008).Human cell suspensions without stimulation agents known to induce CCL5production were used as a baseline control for the experiments. Cellcultures were treated with various exemplar compounds according toFormula 1 at several concentrations for 6 hours prior to stimulation byCCL5 inducing agents. The cultures of NHEK, NHBE, and HUVEC were thenstimulated with various known stimulators of CCL5 production thatconsisted of: Poly [I:C] 10 ug/ml (see FIG. 1a and FIG. 2a ), Flagellin10 ug/ml (Invivogen, San Diego, Calif.; see FIG. 1b ) and TNF alpha 1μg/ml (Invivogen, San Diego, Calif.; see FIG. 2b , FIG. 3, and FIG. 4)for 24-48 hrs). Measurement of CCL5 gene expression was determined fromtotal RNA extracted from cell pellets after incubation by RNAseqquantitation method standardized using expression of housekeeping genesas reference standard. CCL5 protein production was measured in the cellculture supernatants after incubation using the CCL5 ELISA kit (R&DSystems, Minneapolis, Minn.).

To determine the ability of compounds according to Formula 1 to inhibitthe biological effect of CCL5 to induce the transcription of genes andproduction of corresponding proteins known to be upregulated by CCL5 onbinding to its cellular receptors using a functional assay, freshlyisolated human PBMCs or CD14+ monocytes from healthy volunteers wereisolated and cultured at 1×106 cells/ml in RPMI-1640 medium (GIBCO® Inc.Carlsbad, Calif., USA) supplemented with 20% fetal bovine serum and 1%streptomycin/penicillin. The cultures of PBMC, CD+14 monocytes werestimulated with 300 ng/ml of human CCL5 (R&D SYSTEMS®, Minneapolis,Minn. USA) for 4 hrs. Cell suspensions without CCL5 stimulation wereused as a baseline control for the experiments. Cell cultures weretreated with various exemplar compounds according to Formula 1 atseveral concentrations. As a known positive control for CCL5 pathwayinhibition, a blocking antibody to human CCL5 (R&D SYSTEMS®,Minneapolis, Minn. USA) was used (10 ug/ml) that prevents binding ofCCL5 to its receptors, thereby blocking signal transduction through theCCL5 signaling pathways thus inhibiting its biological activity. After24 hours total RNA was extracted from cell pellets and mRNA wasquantitated using expression of housekeeping genes as referencestandard. IL-8 and MMP-19 protein production was measured in the cellculture supernatants after incubation using ELISA (R&D Systems,Minneapolis, Minn.).

To test the effect of the compounds on treatment of human dermatologicdisorders associated with increased CCL5 production by keratinocytes,Compound 11 and Compound 21 were formulated into a topically appliedcream at a concentration of 7%. The compounds in the cream base wereapplied topically to human subjects suffering from dermatitis in an areaof the skin with active skin lesions. The control was the topical creambase composition without the active compounds. Outcome was measured byreduction or disappearance to the lesion in the treated area vs thecontrol area.

Hidradenitis: Three patients with discrete lesions of hidradenitis weretreated with topically applied Compound 20 twice per day for a period of21 days. An additional 4 patients were treated with Compound 16 in asimilar fashion and 3 others with Compound 97 in the same way. In alltreatment groups the lesions treated with the compounds in the creambase decreased and resolved by the 28th day, whereas the lesions on thesame patients treated with the cream base control did not resolve.

Eczema: Four patients with hand and facial eczema were treated withCompound 97 topically applied in a cream base over a period of 14 days.By day 7 the eczema had resolved in the area treated in three of thepatients and by day 12 in the fourth patient. The eczema persisted inthe areas not so treated.

Rosacea: Two patients with rosacea were treated with Compound 25 for 28days in a lotion form. At the end of the course of therapy the rosaceahad nearly disappeared in the treated area, but not in the areasuntreated.

To test the effect of the compounds on treatment of human endothelialdisorders such as abnormal angiogenesis associated with increased CCL5levels. Compound 10 and Compound 21 were formulated into a topicallyapplied solution of 1%. The compounds in the solution were appliedtopically to cultivated human endothelial cells induced to form tubesformed in vitro by CCL5. The control was the topical base solutionwithout the active compounds. Outcome was measured by reduction or lackof formation of the endothelial tubes in the endothelial tube formationassay, which is a human model for abnormal angiogenesis(ThermoFisher-InvitroGen Carlsbad Calif. 92008).

The results of the experiments and clinical tests herein demonstratethat compounds according to Formula 1 inhibit the production of CCL5from human epithelial and endothelial cells and tissues. In addition,they demonstrate a clinical therapeutic effect in patients sufferingfrom a dermatologic disorder associated with production of CCL5 fromkeratinocytes and on CCL5 driven angiogenesis. Therefore, the compoundsaccording to Formula 1 are useful in treatment of inflammatory disordersassociated with increased CCL5 production by epithelial and endothelialtissues including but not limited to: dermatitis, eczema, rosacea andabnormal angiogenesis induced by CCL5.

REFERENCES

-   1. Murphy (2002) Pharmacol Rev 54:227-229-   2. Allen et al. (2007) Annu Rev Immunol 25:787-820-   3. Russo et al. (2010) Curr Opin Drug Discov Devel 13:414-27-   4. Murphy (1994) Annu Rev Immunol 12:593-633-   5. Schein et al. (1988) J Immunol 141:1018-25-   6. Zhebrun et al. (2014) Bull Exp Biol Med. 158(2):192-6-   7. Chihara et al. (1997) J Allergy Clin Immunol. 100(6 Pt 2):S52-5-   8. Toebak et al. (2006) Toxicol In Vitro. 20(1):117-24-   9. Tanaka et al. (2006) Int J Immunogenet. 33(6):423-8-   10. Tanaka et al. (2006) Int J Immunogenet. 33(6):423-8-   11. Nickel et al. (2000) J Immunol. 164(3):1612-6-   12. Antinolo et al. (2003) Mol Hum Reprod. 9(8):491-5-   13. Iijima et al. (2003) Am J Pathol. 163(1):261-8-   14. Yamamoto et al. (2013) Dis Markers. 34(3):153-61-   15. Szodoray et al. (2004) Scand J Immunol. 59(6):592-9-   16. Chen et al. (2004) Tissue Antigens 63(1):41-5-   17. Turner et al. (2014) Biochem Biophys ACTA 1843(11): 2563-2582-   18. Lin et al. (2012) Cancer Sci. 103(5):904-12-   19. Mitchell and Olive (2010) Mol Immunol. 47(11-12):2065-73-   20. Murdoch and Finn (2000) Blood 15; 95(10):3032-43-   21. Camargo et al. (2009) J Immunol. 182(1):171-82-   22. Nickel et al. (2000) J Immunol. 164(3):1612-6-   23. Makki et al. (2000) Clin Exp Rheumatol. 18(3):391-3-   24. Marques et al. (2013) Expert Opin Ther Targets 17(12):1439-60

1. A method of treating a disorder or condition of the skin, mucosalepithelium, and/or pulmonary epithelium comprising topicallyadministering to an area of cells exhibiting the disorder or conditionof a mammalian subject in need thereof a therapeutically effectiveamount of a compound of Formula 1

wherein: n and p are independently selected from 0 or 1; R₁ and R₂ areindependently selected from H, OH, F, Cl, Br, I, (halogen)alkyl, C₁-C₈straight or branched chain alkyl, C₁-C₈ cycloalkyl, heterocycloalkyl,alkylheterocycloalkyl, optionally substituted C₁-C₈ alkenyl, optionallysubstituted C₁-C₈ alkynyl, optionally substituted aryl, optionallysubstituted alkylaryl, optionally substituted heteroaryl, optionallysubstituted alkylheteroaryl, O-alkyl, O-cycloalkyl, O-alkylcycloalkyl,O-aryl, O-optionally substituted aryl, alkyl-O-aryl, alkyl-O-optionallysubstituted aryl, C(O)-aryl, C(O)-optionally substituted aryl,CH₂C(O)-aryl, CH₂C(O)-optionally substituted aryl, O-(halogen)alkyl; R₃,R₄, R₅, R₆, R₇, and R₈, if present, are independently selected from H,C₁-C₈ straight or branched chain alkyl, C₁-C₈ cycloalkyl,heterocycloalkyl, alkylheterocycloalkyl, optionally substituted C₁-C₈alkenyl, optionally substituted C₁-C₈ alkynyl, optionally substitutedaryl, optionally substituted alkylaryl, optionally substitutedheteroaryl, or optionally substituted alkylheteroaryl, and/or adjacentsubstituents R₁ and R₂, R₃ and R₄, R₅ and R₆, R₇ and R₈, if present, mayform a saturated or unsaturated 3-7 membered carbocyclic or heterocyclicring; R₉ is selected from COOH, COO-straight or branched chain alkyl, ora pharmaceutically acceptable salt, ester or prodrug form thereof withthe proviso that when n and p are both 0, R₁ is H, R₃ is methyl, R₁ ismethyl, and R₉ is COOH, COOCH₃, or COOC₂H₅; then R₂ cannot be Cl, andwhen n and p are both 0, R₂ is H, R₃ is methyl, R₁ is methyl, and R₉COOH, COOCH₃, or COOC₂H₅, then R₁ cannot be Cl.
 2. The method accordingto claim 1,

wherein: n and p are independently selected from 0 or 1; R₁ and R₂ areindependently selected from H, OH, F, Cl, Br, C₁-C₈ straight or branchedchain alkyl, CF₃, CH₂CF₃, CF₂CF₃, C₁-C₈ cycloalkyl, optionallysubstituted C₁-C₈ alkenyl optionally substituted C₁-C₈ alkynyl,optionally substituted aryl, optionally substituted alkylaryl,optionally substituted heteroaryl, optionally substitutedalkylheteroaryl, O-alkyl, O-cycloalkyl, O-alkylcycloalkyl, O-aryl,O-optionally substituted aryl, alkyl-O-aryl, alkyl-O-optionallysubstituted aryl, C(O)-aryl, C(O)-optionally substituted aryl,CH₂C(O)-aryl, CH₂C(O)-optionally substituted aryl, OCF₃, OCH₂CF₃,OCF₂CF₃; R₃, R₄, R₅, R₇, and R₈, if present, are independently selectedfrom H, C₁-C₈ straight or branched chain alkyl, C₁-C₈ cycloalkyl,heterocycloalkyl, alkylheterocycloalky, optionally substituted C₁-C₈alkenyl, optionally substituted C₁-C₈ alkynyl, optionally substitutedaryl, optionally substituted alkylaryl, optionally substitutedheteroaryl, optionally substituted alkylheteroaryl, and wherein adjacentsubstituents R₁ and R₂, R₃ and R₄, R₅ and R₆, R₇ and R₈, if present, mayform a saturated or unsaturated 3-7 membered carbocyclic or heterocyclicring; R₉ is selected from COOH, COO-straight or branched chain alkyl, ora pharmaceutically acceptable salt, ester or prodrug form thereof withthe proviso that when n and p are both 0, R₁ is H, R₃ is methyl, R₄ ismethyl, and R₉ is COOH, COOCH₃, or COOC₂H₅, then R₂ cannot be C₁, andwhen n and p are both 0, R₂ is H, R₃ is methyl, R₄ is methyl, and R₉COOH COOCH₃, or COOC₂H₅, then R₁ cannot be Cl.
 3. The method accordingto claim 1,

wherein: n and p are independently selected from 0 or 1; R₁ and R₂ areindependently selected from H, F, Br, CH₃, C₂H₅, C₃H₇, CF₃, CH₂CF₃,CF₂CF₃, C₁-C₈ cycloalkyl, optionally substituted C₁-C₈ alkenyl,optionally substituted C₁-C₈ alkynyl optionally substituted aryl,optionally substituted alkylaryl, optionally substituted heteroaryl,optionally substituted alkylheteroaryl, O-alkyl, O-cycloalkyl,O-alkylcycloalkyl, O-aryl, O-optionally substituted aryl, alkyl-O-aryl,alkyl-O-optionally substituted aryl, C(O)-aryl, C(O)-optionallysubstituted aryl, CH₂C(O)-aryl, CH₂C(O)-optionally substituted aryl,OCF₃, OCH₂CF₃, OCF₂CF₃; R₃, R₄, R₅, R₆, R₇, and R₈, if present, areindependently selected from H, C₁-C₈ straight or branched chain alkyl,C₁-C₈ cycloalkyl, heterocycloalkyl, alkylheterocycloalkyl, optionallysubstituted C₁-C₈ alkenyl, optionally substituted C₁-C₈ alkynyl,optionally substituted aryl, optionally substituted alkylaryl,optionally substituted heteroaryl, optionally substitutedalkylheteroaryl and/or adjacent substituents R₁ and R₂, R₃ and R₄, R₅and R₆, R₇ and R₈, if present, may form a saturated or unsaturated 3-7membered carbocyclic or heterocyclic ring; R₉ is selected from COOH,COO-straight or branched chain alkyl, or a pharmaceutically acceptablesalt, ester, or prodrug form thereof with the proviso that when n and pare both 0, R₁ is H, R₃ is methyl, R₄ is methyl, and R₉ is COOH, COOCH₃,or COOC₂H₅, then R₂ cannot be C₁, and when n and p are both 0, R₂ is H,R₃ is methyl, R₄ is methyl, and R₉ COOH, COOCH₃, or COOC₂H₅, then R₁cannot be Cl.
 4. The method according to claim 1, wherein

wherein: n and p are independently selected from 0 or 1; R₁ and R₂ areindependently selected from H, F, Cl, Br, CH₃, C₂H₅, C₃H₇, CF₃, CH₂CF₃,CF₂CF₃, C₁-C₈ cycloalkyl, optionally substituted C₁-C₈ alkenyl,optionally substituted C₁-C₈ alkynyl, optionally substituted aryl,optionally substituted alkylaryl, optionally substituted heteroaryl,optionally substituted alkylheteroaryl, O-alkyl, O-cycloalkyl,O-alkylcycloalkyl, OC₆H₅, CH₂OC₆H₅, CH₂CH₂OC₆H₅, (CH₂)₃OC₆H₅, OC₆H₄C₁,C(O)C₆H₅, C(O)C₆H₄C₁, CH₂C(O)C₆H₄C₁, OCF₃, OCH₂CF₃, OCF₂CF₃; R₃, R₄, R₅,R₆, R₇, and R₈, if present, are independently selected from H, CH₃,C₂H₅, C₃H₇ and/or adjacent substituents R₁ and R₂, R₃ and R₄, R₅ and R₆,R₇ and R₈, if present, may form a saturated or unsaturated 3-7 memberedcarbocyclic or heterocyclic ring; R₉ is selected from COOH, COO-straightor -branched chain alkyl, or a pharmaceutically acceptable salt, esteror prodrug form thereof, with the proviso that when n and p are both 0,R₁ is H, R₃ is methyl, R₄ is methyl, and R₉ is COOH, COOCH₃, or COOC₂H₅,then R₂ cannot be C₁, and when n and p are both 0, R₂ is H, R₃ ismethyl, R₄ is methyl, and R₉ COOH, COOCH₃, or COOC₂H₅, then R₁ cannot beCl.
 5. The method according to claim 1, wherein the compound of Formula1 is any one of compounds 1-642, or a pharmaceutically acceptable salt,ester, or prodrug form thereof. 6.-17. (canceled)
 18. The methodaccording to claim 1, further comprising a second compound selected fromthe group consisting of Non-steroidal anti-inflammatory drugs,Immunomodulatory agents, Anti-malarials, Antibiotics, Anti-TNF alphaagents, Anti-CD20 agents, Anti-diarrheal drugs, Antidepressants,Anti-psychotics, Anti-fungals, Anti-helminthics, T lymphocyte activationinhibitors, Anti-IL-1 agents, Glucocorticoids, Anti-cytokine/chemokinemonoclonal antibodies, Sex steroids and receptor modulators,Anti-cellular surface receptor monoclonal antibodies, Aminosalicylicacid derivatives, Anticholinergic agents, Adrenergic agonists,Corticosteroids, Anti-neoplastic chemotherapeutic agents,Phosphodiesterase inhibitors, Leukotriene pathway modulators, Monoclonalantibodies directed against human immunoglobulins, Adrenergicantagonists, Calcium channel antagonists, Dopamine agonists, Serotoninagonists, Dopamine antagonists, Serotonin antagonists, Monoaminereuptake inhibitors, Protease inhibitors, Histamine antagonists, Protonpump inhibitors, and HMG-CoA reductase inhibitors.
 19. The methodaccording to claim 1, wherein the disorder is Alopecia, Asthma,Dermatitis, Bronchiolitis, Bronchiolitis Obliterans Syndrome, ChronicObstructive Pulmonary Disease, Dry Eye Syndrome, Dermatomyositis,Esophagitis, Colitis, Hidradenitis, Idiopathic Pulmonary Fibrosis,Macular degeneration, Nasal Polyps, Retinopathy, Scleroderma, Sjogren'ssyndrome, and/or Vitiligo.
 20. The method according to claim 1, whereinthe skin disorder is rosacea.
 21. The method according to claim 1,wherein the skin disorder is hidradenitis.
 22. The method according toclaim 1, wherein the skin disorder is eczema.
 23. The method accordingto claim 1, wherein the skin disorder is atopic dermatitis (AD). 24.(canceled)
 25. The method according to claim 1, wherein the cells arefreshly isolated human cells.
 26. The method according to claim 25,wherein the human cells are keratinocytes.
 27. The method according toclaim 25, wherein the human cells are leukocytes.
 28. (canceled)
 29. Amethod of inhibiting the biological activity of CCL5 on mammalian cellscomprising topically administering to a mammalian subject in needthereof a therapeutically effective amount of a compound according toFormula 1 or a tautomer thereof.

wherein: n and p are independently selected from 0 or 1; Re and R₂ areindependently selected from H, OH, F, Cl, Br, I, (halogen)alkyl, C₁-C₈straight or branched chain alkyl, C₁-C₈ cycloalkyl, heterocycloalkyl,alkylheterocycloalkyl, optionally substituted C₁-C₈ alkenyl, optionallysubstituted C₁-C₈ alkynyl, optionally substituted aryl, optionallysubstituted alkylaryl, optionally substituted heteroaryl, optionallysubstituted alkylheteroaryl, O-alkyl, O-cycloalkyl, O-alkylcycloalkyl,O-aryl, O-optionally substituted aryl, alkyl-O-aryl, alkyl-O-optionallysubstituted aryl, C(O)-aryl, C(O)-optionally substituted aryl,CH₂C(O)-aryl, CH₂C(O)-optionally substituted aryl, O-(halogen)alkyl; R₃,R₄, R₅, R₆, R₇, and R₈, if present, are independently selected from H,C₁-C₈ straight or branched chain alkyl, C₁-C₈ cycloalkyl,heterocycloalkyl, alkylheterocycloalkyl, optionally substituted C₁-C₈alkenyl, optionally substituted C₁-C₈ alkynyl, optionally substitutedaryl, optionally substituted alkylaryl, optionally substitutedheteroaryl, or optionally substituted alkylheteroaryl, and/or adjacentsubstituents R₁ and R₂, R₃ and R₄, R₅ and R₆, R₇ and R₈, if present, mayform a saturated or unsaturated 3-7 membered carbocyclic or heterocyclicring; R₉ is selected from COOH, COO-straight or branched chain alkyl, ora pharmaceutically acceptable salt, ester or prodrug form thereof withthe proviso that when n and p are both 0, R₁ is H, R₃ is methyl, R₄ ismethyl, and R₉ is COOH, COOCH₃, or COOC₂H₅ then R₂ cannot be Cl, andwhen n and p are both 0, R₂ is H, R₃ is methyl, R₄ is methyl, and R₉COOH, COOCH₃, or COOC₂H₅, then Re cannot be Cl.
 30. A method of treatinga disease or condition of the skin, mucosal epithelium, pulmonary, orgastrointestinal tract epithelium associated with increased levels ofCCL5 by topically administering to an area of cells exhibiting thedisorder or condition of a mammalian subject in need thereof atherapeutically effective amount of a compound of Formula 1

wherein: n and p are independently selected from 0 or 1; R₁ and R₂ areindependently selected from H, OH, F, Cl, Br, I, (halogen)alkyl, C₁-C₈straight or branched chain alkyl, C₁-C₈ cycloalkyl, heterocycloalkyl,alkylheterocycloalkyl, optionally substituted C₁-C₈ alkenyl, optionallysubstituted C₁-C₈ alkynyl, optionally substituted aryl, optionallysubstituted alkylaryl, optionally substituted heteroaryl, optionallysubstituted alkylheteroaryl, O-alkyl, O-cycloalkyl, alkylcycloalkyl,O-aryl, O-optionally substituted aryl, alkyl-O-aryl, alkyl-O-optionallysubstituted aryl, C(O)-aryl, C(O)-optionally substituted aryl,CH₂C(O)-aryl, CH₂C(O)-optionally substituted aryl, O-(halogen)alkyl; R₃,R₄, R₅, R₆, R₇, and R₈, if present, are independently selected from H,C₁-C₈ straight or branched chain alkyl, C₁-C₈ cycloalkyl,heterocycloalkyl, alkylheterocycloalkyl, optionally substituted C₁-C₈alkenyl, optionally substituted C₁-C₈ alkynyl, optionally substitutedaryl, optionally substituted alkylaryl, optionally substitutedheteroaryl, or optionally substituted alkylheteroaryl, and/or adjacentsubstituents Re and R₂, R₃ and R₄, R₅ and R₆, R₇ and R₈, if present, mayform a saturated or unsaturated 3-7 membered carbocyclic or heterocyclicring; R₉ is selected from COOH, COO-straight or branched chain alkyl, ora pharmaceutically acceptable salt, ester or prodrug form thereof withthe proviso that when n and p are both 0, R₁ is H, R₃ is methyl, R₄ ismethyl, and R₉ is COOH, COOCH₃ or COOC₂H₅, then R₂ cannot be Cl, andwhen n and p are both 0, R₂ is H, R₃ is methyl, R₄ is methyl, and R₉COOH, COOCH₃, or COOC₂H₅, then R₁ cannot be C₁.
 31. The method accordingto claim 29, further comprising administering at least one othercompound selected from the group consisting of: Non-steroidalanti-inflammatory drugs, Immunomodulatory agents, Anti-malarials,Antibiotics, Anti-TNF alpha agents, Anti-CD20 agents, Anti-diarrhealdrugs, Antidepressants, Anti-psychotics, Anti-fungals, Anti-helminthics,T lymphocyte activation inhibitors, Anti-IL-1 agents, Glucocorticoids,Anti-cytokine/chemokine monoclonal antibodies, Sex steroids and receptormodulators, Anti-cellular surface receptor monoclonal antibodies,Aminosalicylic acid derivatives, Anticholinergic agents, Adrenergicagonists, Corticosteroids, Anti-neoplastic chemotherapeutic agents,Glucose modulators such as insulin, glyburide, and metformin,Phosphodiesterase inhibitors, Leukotriene pathway modulators, Monoclonalantibodies directed against human immunoglobulins, Adrenergicantagonists, Calcium channel antagonists, Dopamine agonists, Serotoninagonists, Dopamine antagonists, Serotonin antagonists, Monoaminereuptake inhibitors, Protease inhibitors, Histamine antagonists, Protonpump inhibitors, and HMG-CoA reductase inhibitors.
 32. The methodaccording to claim 29, wherein the cells are freshly isolated humancells.
 33. The method according to claim 1, wherein the cells areprimary culture human cells.
 34. The method according to claim 29,wherein the cells are primary culture human cells.